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PMID: 11013226 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Ribosomal protein L2 is involved in the association of the ribosomal subunits, tRNA binding to A and P sites and peptidyl transfer.

The EMBO journal ·Vol. 19 ·No. 19 ·2000-10-02 ·Pages 5241-50

Diedrich G, Spahn CM, Stelzl U, Schäfer MA, Wooten T, Bochkariov DE, Cooperman BS, Traut RR, Nierhaus KH

Abstract

Ribosomal proteins L2, L3 and L4, together with the 23S RNA, are the main candidates for catalyzing peptide bond formation on the 50S subunit. That L2 is evolutionarily highly conserved led us to perform a thorough functional analysis with reconstituted 50S particles either lacking L2 or harboring a mutated L2. L2 does not play a dominant role in the assembly of the 50S subunit or in the fixation of the 3'-ends of the tRNAs at the peptidyl-transferase center. However, it is absolutely required for the association of 30S and 50S subunits and is strongly involved in tRNA binding to both A and P sites, possibly at the elbow region of the tRNAs. Furthermore, while the conserved histidyl residue 229 is extremely important for peptidyl-transferase activity, it is apparently not involved in other measured functions. None of the other mutagenized amino acids (H14, D83, S177, D228, H231) showed this strong and exclusive participation in peptide bond formation. These results are used to examine critically the proposed direct involvement of His229 in catalysis of peptide synthesis.

MeSH Terms
Amino Acid Sequence Binding Sites Catalytic Domain Escherichia coli/genetics,metabolism Histidine/chemistry,metabolism Molecular Sequence Data Mutation Peptidyl Transferases/metabolism Protein Biosynthesis RNA, Transfer/chemistry,metabolism Ribosomal Proteins/chemistry,genetics,metabolism Ribosomes/chemistry,genetics,metabolism Sequence Alignment
Chemicals
Ribosomal Proteins ribosomal protein L2 Histidine RNA, Transfer Peptidyl Transferases
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Diedrich G
Max-Planck-Institut für Molekulare Genetik, AG Ribosomen, Ihnestrasse 73, D-14195 Berlin, Germany.
Spahn C M
Stelzl U
Schäfer M A
Wooten T
Bochkariov D E
Cooperman B S
Traut R R
Nierhaus K H
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/19.19.5241
Published
2000-10-02
Pages
5241-50
Language
English
Region
England
NLM ID
8208664
PMCID
PMC302109
Subset
IM
Grants
NIGMS NIH HHS · F32 GM017924 · United States
NIGMS NIH HHS · GM 17924 · United States
NIGMS NIH HHS · GM 53146 · United States
External Links
PubMed source DOI: 10.1093/emboj/19.19.5241 PMC Full Text View journal details
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